EP0864090A1 - Procede et appareil destines a l'evaluation quantitative et semi-quantitative d'une substance a analyser - Google Patents

Procede et appareil destines a l'evaluation quantitative et semi-quantitative d'une substance a analyser

Info

Publication number
EP0864090A1
EP0864090A1 EP96928285A EP96928285A EP0864090A1 EP 0864090 A1 EP0864090 A1 EP 0864090A1 EP 96928285 A EP96928285 A EP 96928285A EP 96928285 A EP96928285 A EP 96928285A EP 0864090 A1 EP0864090 A1 EP 0864090A1
Authority
EP
European Patent Office
Prior art keywords
calibration
zone
analyte
lateral flow
agent
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP96928285A
Other languages
German (de)
English (en)
Other versions
EP0864090A4 (fr
Inventor
Dennis Brian Rylatt
Dean Moss
Andrew Jane
Peter Gregory Bundesen
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Agen Biomedical Ltd
Original Assignee
Agen Biomedical Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Agen Biomedical Ltd filed Critical Agen Biomedical Ltd
Publication of EP0864090A1 publication Critical patent/EP0864090A1/fr
Publication of EP0864090A4 publication Critical patent/EP0864090A4/fr
Withdrawn legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody

Definitions

  • THIS INVENTION relates generally to analyte assays.
  • this invention relates to an assay and apparatus for semi- quantitative determination of an analyte in a test sample.
  • the invention also relates to quantitative determination of an analyte by use of an apparatus which reads and processes the results of the assay of the invention. BACKGROUND OF THE INVENTION
  • U.S. Patent Specification 5,073,484 is directed to a method and an apparatus for the quantification of an analyte in a liquid employing a liquid- permeable solid medium defining a liquid flow path.
  • the medium includes a number of reaction zones, containing a reactant, spaced apart along the flow path and in which reaction occurs between the reactant and the analyte or an analyte derivative (e.g. a labelled analyte) to result in the formation of a predetermined product.
  • Detector means are employed to detect analyte, analyte derivatives, reactant or predetermined product in the reaction zones, the number of such zones in which such detection occurs indicating the amount of analyte in the liquid.
  • the method is predicated on the condition that for a given amount of analyte in a liquid, the analyte must be bound substantially to saturation in a first reaction zone before any residual amount of the analyte is bound subsequently in a second reaction zone and so on.
  • this method provides theoretically for a linear relationship between the number of reaction zones in which detection occurs and the amount of analyte in a liquid.
  • US Patent 5,073,484 in including a multiplicity of reaction zones does not include a calibration zone which has a calibration agent receptor bound thereto.
  • a further attempt at developing a semi-quantitative assay is disclosed in Australian Patent Specification 658566.
  • One embodiment of this assay utilizes a vertical flow-through filter comprising a test zone and one or more calibration zones. At the test zone there is non-diffusibly attached therein an analyte receptor capable of binding a target analyte, and at the or each calibration zone there is non-diffusibly attached therein a calibration receptor capable of binding a calibration agent. When there is more than one calibration zone, one of these zones is capable of binding a higher predetermined amount of the calibration agent than the other.
  • a test sample suspected of containing the target analyte is introduced to the filter and the target analyte, if present, binds to the analyte receptor at the test zone.
  • a solution comprising a labeled analyte detection agent as well as a labeled calibration agent is added subsequently to the filter.
  • the analyte detection agent binds to the target analyte at the test zone and the calibration agent binds to the calibration receptor at the calibration zone in amounts which are directly proportional to the amounts of target analyte and calibration receptor bound to the filter.
  • the presence of analyte detection agent bound at the test zone and the presence of calibration agent bound at the or each calibration zone is determined subsequently by signal development, permitting qualitative or semi-quantitative determination of the target analyte in the test sample.
  • the target analyte in the test sample is premixed with a solution comprising labeled analyte detection agent and labeled calibration agent prior to introduction of the mixture onto the filter.
  • a method for quantitative or semi-quantitative determination of target analyte(s) in a test sample comprising the steps of:
  • the target analyte(s) may be selected from a group including an antigenic substance, a hapten, an antibody, a protein, a peptide, an amino acid, a nucleic acid, a hormone, a steroid, a vitamin, a carbohydrate, a lipid, a blood clotting factor, a pathogenic organism for which polyclonal and/or monoclonal antibodies can be produced, a natural or synthetic chemical substance, a contaminant, a drug including those administered for therapeutic purposes as well as those administered for illicit purposes, and metabolites of or antibodies to any of the above substances.
  • test zones utilized in the lateral flow liquid permeable medium will be dependent on the number of target analytes being quantified. In the preferred embodiment, however, the lateral flow liquid permeable medium will only include a single test zone for quantification of a single target analyte.
  • the target analyte has two binding sites each of which is capable of forming a specific binding pair with a specific binding partner.
  • a specific binding pair comprises two different molecules wherein one ofthe molecules through chemical or physical means specifically binds to a second molecule.
  • specific binding partners examples of which are described in U.S. Patent Specification 5,075,078, include antigens and antibodies, lectins and carbohydrates, complementary peptides, protein, carbohydrate and nucleic acid structures, enzyme inhibitors and enzymes, Protein A and IgG as well as effector and receptor molecules. It will of course be appreciated that it is sufficient for the target analyte to have a single binding site for a specific binding partner if the target analyte in combination with a first specific binding partner is capable of producing a unique binding site for a second specific binding partner.
  • test sample is preferably an aqueous solution.
  • test sample as used herein may include a biological fluid which may be extracted, untreated, treated, diluted or concentrated from a plant or animal.
  • the biological fluid is derived from an animal.
  • the biological fluid is selected from a group including whole blood, serum, plasma, saliva, urine, sweat, ascitic fluid, peritoneal fluid, synovial fluid, amniotic fluid, cerebrospinal fluid and the like.
  • the calibration agent and the analyte detection agent are diffusibly attached to the separate support element rather than the lateral flow liquid permeable medium.
  • the separate support element is in fluid communication with the lateral flow liquid permeable medium such that the test sample may pass or migrate from the separate support element to the lateral flow liquid permeable medium.
  • Fluid communication may include physical contact ofthe separate support element to the lateral flow liquid permeable medium as well as the separation of the lateral flow liquid permeable medium by an intervening space or additional material which still allows fluid communication between the element and the liquid permeable medium.
  • the separate support element partially overlaps and is in close proximity to an end portion of the lateral flow liquid permeable medium.
  • the separate support element may form part of one end of the lateral flow liquid permeable medium.
  • the separate support element may be composed of any suitable material which can transfer the test sample to the lateral flow liquid permeable medium.
  • the separate support element is adapted to absorb a volume of test sample which is equal to or greater than the total volume capacity of the lateral flow liquid permeable medium.
  • a material suitable for use as the separate support element may include nitrocellulose, cellulosic paper, porous polyethylene frit or pads and glass fibre filter paper.
  • the separate support element may contain one or more assay reagents either diffusibly or non-diffusibly attached therein.
  • the assay reagents in the separate support element are diffusibly attached therein.
  • Suitable assay reagents that may be contained in the separate support element include the analyte detection agent and the calibration agent.
  • the analyte detection agent is a specific binding partner of the target analyte which may have an associated label.
  • the analyte detection agent may be a predetermined amount of the target analyte itself having a label associated therewith.
  • the analyte which is preferably directly labeled, acts as the analyte detection agent by competing with the analyte in the test sample for binding to a specific binding partner which is non-diffusibly attached in the test zone.
  • an analog ofthe analyte may also be used as the analyte detection agent.
  • analyte detection agent and the calibration agent each have a label associated therewith which includes the following: i. direct attachment of the label to the analyte detection agent; ii. indirect attachment of the label to the analyte detection agent; i.e., attachment of the label to another assay reagent which subsequently binds to the analyte detection agent; and iii. attachment to a subsequent reaction product of the analyte detection agent.
  • the calibration agent has the same label associated therewith which also may include i., ii., and iii. above.
  • the label is attached directly to the analyte detection agent and the calibration agent.
  • the label may be selected from a group including a chromogen, a catalyst, an enzyme, a fluorophore, a chemiluminescent molecule, a lanthanide ion such as Europium (Eu 34 ), a radioisotope and a direct visual label.
  • a colloidal metallic or non-metallic particle a dye particle, an enzyme or a substrate, an organic polymer, a latex particle, a liposome, or other vesicle containing a signal producing substance and the like.
  • a large number of enzymes suitable for use as labels is disclosed in United States Patent Specifications U.S. 4,366,241 , U.S. 4,843,000, and U.S. 4,849,338, all of which are herein incorporated by reference.
  • Suitable enzyme labels useful in the present invention include alkaline phosphatase, horseradish peroxidase, luciferase, ⁇ -galactosidase, glucose oxidase, lysozyme, malate dehydrogenase and the like.
  • the enzyme label may be used alone or in combination with a second enzyme which is in solution.
  • the fluorophore is selected from a group including fluorescein isothiocyanate (FITC), tetramethylrhodamine isothiocyanate (TRITL) or R-Phycoerythrin (RPE).
  • FITC fluorescein isothiocyanate
  • TTL tetramethylrhodamine isothiocyanate
  • RPE R-Phycoerythrin
  • the label is a colloidal metallic particle.
  • the label is colloidal gold.
  • the calibration agent is capable of forming a specific binding pair with a specific binding partner.
  • the calibration agent has a high affinity for the specific binding partner thereof.
  • the calibration agent is most suitably avidin, streptavidin or biotin or other calibration agent which has high affinity for the specific binding partner thereof.
  • the passage of the mixture through the lateral flow liquid permeable medium may be effected by capillary migration, gravity flow or electrophoresis or a medium of hydrophilic nature having an affinity for water.
  • the passage of the mixture is effected by capillary migration.
  • the lateral flow liquid permeable medium may be any suitable absorbent, porous, capillarity-possessing or bibulous material through which a solution containing the target analyte can be transported.
  • Natural, synthetic or naturally occurring materials which are synthetically modified may be used as the liquid permeable medium including but not limited to cellulose materials such as paper, cellulose and cellulose derivatives such as cellulose acetate and nitrocellulose; glass fibres; natural or synthetic cloth; porous gels such as agarose, silica gel, dextran and gelatin; porous fibrous matrixes; starch based materials such as Sephadex® cross-linked dextran chains; ceramic materials; films of polyvinyl chloride and polyamide; and combinations of polyvinyl chloride-silica and the like.
  • cellulose materials such as paper, cellulose and cellulose derivatives such as cellulose acetate and nitrocellulose
  • glass fibres such as cellulose acetate and nitrocellulose
  • porous gels such as agarose, silica gel, dextran and gelatin
  • porous fibrous matrixes such as starch based materials such as Sephadex® cross-linked dextran chains
  • ceramic materials films of polyvinyl chloride and polyamide; and
  • the lateral flow liquid permeable medium is a cellulose material.
  • the lateral flow liquid permeable medium is nitrocellulose.
  • the lateral flow liquid permeable medium does not interfere with the production of the detectable signal.
  • the method of the invention may also include an additional step of filtering the test sample.
  • the filtering may be effected by any filter or trapping device used to remove particles above a certain size from the test sample.
  • the filtering step may be used to remove red blood cells from a sample of whole blood, such that plasma is the fluid which is received by the separate support element and transferred to the lateral flow liquid permeable medium.
  • the specific binding partner of the analyte receptor is the analyte or analog thereof.
  • the specific binding partner ofthe analyte receptor may be the analyte detection agent.
  • the analyte detection agent may be a predetermined amount of analyte having a label associated therewith.
  • the at least one calibration zone comprises a predetermined amount of a calibration agent receptor non-diffusibly attached therein which is capable of participating in the formation of an immobilized specific binding pair with the calibration agent.
  • the predetermined amount of the calibration agent receptor is selected so that a substantially finite amount ofthe calibration agent is bound in the at least one calibration zone.
  • the predetermined amount is such that when the substantially finite amount of the calibration agent is bound in the calibration zone, the signal generated therein is within a dynamic range suitable for comparing with the signal generated in the test zone.
  • each calibration zone has a different predetermined amount of the calibration agent receptor so that, in operation, each calibration zone produces a different signal.
  • Non-diffusive attachment of the analyte receptor and the calibration agent receptor may be effected by any suitable means including covalent bonding, non-covalent bonding or entrapment. It will of course be appreciated that the receptors used in the invention may be non-diffusibly attached either directly or indirectly. For example, each receptor may be covalently bound to a spacer molecule which has been covalently bound to the lateral flow liquid permeable medium.
  • the spacer molecule may include a latex microparticle, a protein such as bovine serum albumin (BSA) or a polymer such as dextran.
  • BSA bovine serum albumin
  • dextran a polymer such as dextran
  • the signals generated in the test zone and the at least one calibration zone may be measured by visual inspection or by an instrumental means.
  • the signals are measured by the instrumental means.
  • the signal is preferably a colored signal and the preferred method of measurement is reflectance.
  • Measurement of the signals may be made by any instrument which is capable of converting signal into a digital expression that is proportional to the signal produced.
  • a signal may be instrumentally detected by irradiating a fluorescent label with light and measuring the level of fluorescence in a fluorimeter; by providing for an enzyme system to produce a dye which could be measured using a spectrophotometer; or detection of a dye particle or a colored colloidal metallic or non metallic particle using a reflectometer; in the case of using a radioactive label or chemiluminescent molecule employing a radiation counter.
  • comparison of the signals generated in at least one test zone and the at least one calibration zone is used to determine the approximate concentration of the analyte in the test sample.
  • This comparison is predicated on the condition that a signal correlates with a certain amount or concentration of a label captured in a zone which in turn corresponds proportionally to a specific amount of the analyte.
  • a predetermined amount of a calibration agent receptor in a calibration zone is chosen to bind substantially a finite amount of a calibration agent which in turn must be bound by a certain amount of the label and the signal generated therefrom must be reflective of a specific amount of the analyte.
  • the method of the invention further includes within its scope the use of one or more calibration agents in step (iii) wherein there is also provided one or more complementary calibration agent receptors each of which is non-diffusibly attached respectively in different calibration zones.
  • the method of the invention still further includes within its scope the use of a procedural control step which indicates validation of the methods of the invention.
  • the procedural control step may be effected by inclusion of a procedural control zone which may form part of the lateral flow liquid permeable medium.
  • the completion ofthe method may be indicated by a changing of color which may be effected by an assay reagent contacting the procedural control zone.
  • the assay reagent is the analyte detection agent having a label associated therewith.
  • the assay reagent may be a compound which changes color upon contact with a test solution containing water which compound includes a dehydrated transition metal salt such as CuS0 4 ,
  • a method for quantitative or semi-quantitative determination of target analyte(s) in a test sample comprising the steps of:
  • a support medium which may comprise the lateral flow liquid permeable medium or a separate support element an analyte detection agent which detects the target analyte if present in the test sample, said analyte detection agent having a label associated therewith;
  • a support medium which may comprise the lateral flow liquid permeable medium or a separate support element a calibration agent having the same label associated therewith;
  • the analyte detection agent will comprise a specific binding partner of the target analyte which may have the same label associated therewith as the calibration agent.
  • a lateral flow liquid permeable medium adapted to convey fluids at least one fluid flow direction for use in the method according to the first aspect of the invention comprising:
  • an assay reagent zone including a calibration agent having a label associated therewith and the analyte detection agent which has the same label associated therewith.
  • a lateral flow liquid permeable medium adapted to convey fluids in at least one fluid flow direction for use in the method according to the second aspect of the invention comprising:
  • test zone having non-diffusibly attached therein a predetermined amount of analyte which is capable of participating in the formation of an immobilized specific binding pair with an analyte detection agent
  • an assay reagent zone including a calibration agent having a label associated therewith and the analyte detection agent which has the same label associated therewith.
  • the lateral flow liquid permeable mediums described above further include a separate support element which has diffusibly attached thereon the calibration agent and associated label and the analyte detection agent and associated label.
  • the lateral flow liquid permeable medium preferably includes a filter in fluid communication with said separate support element.
  • the lateral flow liquid permeable medium includes a procedural control zone as described above.
  • the liquid permeable medium may also include an absorbent pad which is downstream of said test and said at least one calibration zone, which absorbent pad is adapted to absorb an excess amount of a test sample or a reagent solution.
  • the liquid permeable medium may include in one situation a forward flow and reverse flow of sample as well as diffusible reagents. However, most preferrably the liquid permeable medium is designed to accommodate only a single fluid flow direction.
  • the invention also comprises a kit for use in the methods as herein described including the lateral flow liquid permeable medium as described above.
  • kit may inco ⁇ orate a housing which inco ⁇ orates the lateral flow liquid permeable medium and optionally the separate support medium as described above with appropriate windows for viewing the test zone and calibration zone and optionally a window for viewing the procedural control zone described above.
  • the housing may also include an aperture for introduction of the test sample suspected of containing the target analyte.
  • the invention may also include a signal measuring device which includes the housing as described above.
  • the signal measuring device measures the signals as described above and is programmed to contain a multiplicity of standard curves for the various analytes which may be obtained from the calibration zones as for example described in an article by Diebold E, et al, Chemtech 1991 ; 462-7 and an article by Burke A, ef al, Chemtech 1991 ; 547-551 which are hereby incorporated by reference.
  • a lysis solution may also be provided suitable for lysis of red blood cells in the test sample.
  • FIG. 1 illustrates the results of two analyte assays utilizing lateral flow membranes constructed in accordance with U.S. 5,073,484 and colloidal gold as the detector means wherein the top illustration refers to a an assay result in which a relatively low amount of target analyte was assayed and the bottom illustration refers to an assay result in which a relatively high amount of target analyte was assayed;
  • FIG. 2 refers to a side view illustration of one embodiment of an unused lateral flow liquid permeable medium constructed in accordance with the invention.
  • FIG. 3 refers to a top plan view of five lateral flow liquid permeable mediums constructed in accordance with FIG. 2 wherein each is contained within a cartridge and wherein different assay results are shown.
  • FIG. 4 refers to a side view illustration of the lateral flow liquid permeable medium of FIG. 2 after completion of an assay in which no target analyte was present in the test sample.
  • FIG. 5 refers to a side view illustration of the lateral flow liquid permeable medium of FIG. 2 after completion of an assay in which target analyte was present in the test sample.
  • FIG. 6 refers to a box plot illustrating a comparison ofthe visual semi-quantitative determination of D-dimer with a quantitative determination of D-dimer by EIA.
  • FIG.7 refers to a box plot illustrating a comparison ofthe visual semi-quantitative determination of myoglobin with a quantitative determination of myoglobin by EIA.
  • FIG. 8 refers to a side view illustration of an alternative embodiment of an unused lateral flow liquid permeable medium constructed in accordance with the invention.
  • FIG. 9 refers to a top plan view of four lateral flow liquid permeable mediums constructed in accordance with FIG. 8 wherein each is contained within a cartridge and wherein different assay results are shown.
  • FIG. 10 refers to a side view illustration ofthe lateral flow liquid permeable medium of FIG. 8 after completion of an assay in which no target analyte was present in the test sample.
  • FIG. 11 refers to a side view illustration of the lateral flow liquid permeable medium of FIG. 8 after completion of an assay in which no target analyte was present in the test sample.
  • FIG. 12 refers to a linear regression analysis corresponding to a comparison of quantitative determination of D-dimer by instrument with quantitative determination of D-dimer by EIA.
  • FIG. 13 refers to a linear regression analysis corresponding to a comparison of quantitative determination of myoglobin by instrument with quantitative determination of myoglobin by EIA.
  • FIG.14 refers to a standard curve of reflectance signal as a function of biotin - BSA concentration.
  • FIG. 2 A side view diagrammatic representation of a device comprising the lateral flow liquid permeable medium constructed in accordance with the invention is shown in FIG. 2.
  • the device consists of a filter 201 adapted to remove red blood cells from a test sample, a nitrocellulose membrane 207 and a separate support element 202 containing a suitable concentration of reagents including the D-dimer binding monoclonal antibody DD-3B6/22 labeled with colloidal gold (3B6-gold; i.e., analyte detection agent) 208 specific for the thrombosis marker D-dimer 213 (i.e., target analyte) under test and streptavidin labeled with colloidal gold (streptavidin-gold; i.e., calibration agent) 209.
  • DD-3B6/22 labeled with colloidal gold
  • streptavidin-gold i.e., calibration agent
  • the separate support element 202 receives the test sample, and the wetting of the element by the test sample performs at least two functions. Firstly, it dissolves or reconstitutes a predetermined amount of 3B6-gold 208 and streptavidin-gold 209. Secondly, it initiates the transfer of both the test sample containing D-dimer 213 and the freshly dissolved reagents 208, 209. Both the 3B6-gold and the streptavidin-gold are mobilized when moistened by the test sample and/or an application buffer.
  • the nitrocellulose membrane 207 comprises a test zone 204 and two calibration zones 210, 211.
  • Non-diffusibly attached at a first calibration zone 210 is a first predetermined amount of BSA labeled with biotin (BSA-biotin; i.e., calibration agent receptor) 214 capable of binding specifically to the streptavidin-gold.
  • BSA-biotin biotin
  • BSA-biotin i.e., calibration agent receptor
  • the concentration of BSA-biotin at the second calibration zone 211 is adjusted to generate a different signal intensity compared to that generated at the first calibration zone 210.
  • DD-1D2/48 1D2 antibody a monoclonal antibody termed DD-1D2/48 1D2 antibody; (i.e., analyte receptor) 215 capable of binding D-dimer when complexed to the 3B6-gold.
  • a procedural control zone 212 containing an anti-mouse antibody 217 which is non-diffusibly attached therein and is capable of specifically binding the 3B6-gold.
  • an absorbent pad 206 At the end of the membrane 207 there is provided an absorbent pad 206 to absorb an excess of test sample, application buffer or assay reagents.
  • the filter 201 which is adapted to remove red blood cells or particulate matter may not be necessary.
  • test sample and/or sample application buffer causes moistening of the separate support element which subsequently mobilizes the 3B6-gold 208 and streptavidin-gold 209.
  • D- dimer 213 If D- dimer 213 is present in the test sample, it binds the 3B6-gold 208 via a first antigenic site to form a complex.
  • This complex in concert with the streptavidin-gold 209, subsequently migrates to the first calibration zone 210 whereat a substantially finite amount ofthe streptavidin-gold binds to the first predetermined amount of BSA-biotin 214 in the first calibration zone 210.
  • the analyte-3B6-gold complex subsequently migrates to the test zone 204 whereat the complex binds the 1 D2 antibody 215 via a second antigenic site of D-dimer 213.
  • Streptavidin-gold 209 which has not bound to the first calibration zone 210 subsequently migrates to the second calibration zone 211 whereat it binds to a second predetermined amount of BSA-biotin 216.
  • the concentrations of the streptavidin-gold 209 and the BSA- biotin 214, 216 are adjusted by methods described in Example 6 to give signal intensities that fall within a suitable dynamic range of those produced by typical diagnostic concentrations of D-dimer in blood or plasma to generate calibration zones which, when in operation, generate signal intensities corresponding to 120 ng/mL and 1000 ng/mL of D-dimer.
  • the introduction of such calibration agents enable visual inspection of a test device to assign one of three values to any test sample (FIG.3).
  • the device as illustrated in FIG. 2 is now contained within a housing or cartridge comprising a sample receiving aperture (S) which is in communication with the separate support element.
  • the housing also includes a window which facilitates viewing ofthe procedural control (C), test (T), as well as the 120 ng/mL and
  • FIG. 3a there is shown a top view of an unused device for semi-quantitative measurement of human D-dimer. Note that there are no visible or instrumental read lines in the viewing window.
  • the test device After the successful completion of a negative test. Note the appearance of zones of color in the viewing window at the 120 ng/mL and 1000 ng/mL calibration zones and the procedural control zone and the absence of any color at the test zone. This allows a value of less than 120 ng/mL of D-dimer to be assigned to the test sample.
  • FIG. 3c there is shown the device after the successful completion of a test with a sample containing less than 120 ng/mL D-dimer.
  • a sample containing less than 120 ng/mL D-dimer there is a visible line at the test zone but of lower intensity than the 120 ng/mL calibration zone thus allowing a value of less 120 ng/mL of D-dimer to be assigned to this test sample.
  • the respective intensities of the other zones are similar to those in FIG. 3b.
  • FIG. 3d there is shown the device after the successful completion of a test on a sample containing a D-dimer concentration of between 120 ng/mL and 1000 ng/mL. Note the zone of intensity in the test zone is now intermediate between the 120 ng/mL and
  • FIG. 3e shows the device after the successful completion of a test on a sample containing more than 1000 ng/mL D-dimer.
  • FIG. 4 is a side view diagrammatic representation ofthe assay reagents after completion of a test in which no D-dimer was present in the sample.
  • the labeled reagents 208, 209 have migrated from the separate support element 202 and the streptavidin-gold 209 has bound to the calibration zones 210, 211.
  • the 3B6-gold 208 has passed through the test zone 204 without significant binding and consequently, there was no visible or instrument determinable signal at the test zone 204.
  • FIG. 5 is a side view diagrammatic representation of a completed test in which a significant concentration of D-dimer 213 was present in the test sample.
  • the streptavidin-gold 209 not only has the streptavidin-gold 209 been captured at the respective calibration zones 210, 211 but the analyte-3B6-gold complex has also been captured at the test zone 204.
  • An estimation of the concentration of the D-dimer 213 in the test sample is made by comparing the signal intensity generated in the test zone 204 with those generated in the respective calibration zones 210, 211.
  • D-dimer with a quantitative determination of D-dimer by enzyme immunoassay (EIA) (DIMER TEST EIA, Agen Biomedical, Australia) is shown in FIG. 6.
  • EIA enzyme immunoassay
  • the data is presented as a box plot.
  • the visual semi- quantitative determination was made with 187 plasma samples of clinical origin using 25 ⁇ L of sample.
  • the visual intensity of the test zone was compared with the visual intensity of the two calibration zones.
  • the results show that the 102 samples graded visually to be less than the 120 ng/ml had a median value of 46 ng/ml when measured by EIA, 75% were less than 80 ng/ml and 90% were less than 150 ng/ml.
  • test is performed on serum, plasma, anti-coagulated whole blood samples (citrate, EDTA, heparin) or finger stick samples directly applied to the test device using the following procedure.
  • the components are allowed to come to room temperature prior to use, keeping a pouch containing the test devices sealed to avoid condensation on the test surface.
  • the pouch is then opened and the test devices placed on a flat horizontal surface.
  • the device comprises a first calibration zone corresponding to a value of 70 ng/mL myoglobin, a second calibration zone corresponding to a value of 350 ng/mL myoglobin and a test zone therebetween. The test is valid if, after 10 minutes, a color appears in the
  • Samples with myoglobin values less than 70 ng/mL at the test zone (T) have either no color in the test zone or a color of equal intensity or less compared to that in the 70 ng/mL calibration zone.
  • Samples with myoglobin levels higher than 70 ng/mL may be of clinical significance and result in a color in the test zone of intensity greater than that in the 70 ng/mL calibration zone. If the colour in the test zone is less intense than that in the 350 ng/mL calibration zone, then the sample has a value less than 350 ng/mL.
  • FIG. 7 A comparison of the visual semi-quantitative determination of myoglobin with a quantitative determination of myoglobin by EIA (Stratus EIA, DADE, USA) is shown in FIG. 7.
  • the data is presented as a box plot.
  • the visual semi-quantitative determination was made with 127 plasma samples of clinical origin using 25 ⁇ L of sample. At ten minutes after the tests were run the visual intensity of the test zone was compared to the visual intensity of the two calibration zones.
  • the results show that the 59 samples graded visually to be less than the 70 ng/ml had a median value of 47 ng when measured by EIA, 75% were less than 70 ng/ml and 90% were less than 85 ng/ml. Samples graded visually to be between 70 ng/ml and 350 ng/ml usually had values in this range. The higher the myoglobin level in the sample the more likely a high visual grading.
  • the device can also be configured to allow the semi- quantitation of smaller analytes such as drugs, including therapeutic drugs and illicit drugs, hormones, vitamins, steroids, peptides and haptens as well as the larger molecular weight molecules which can be assayed in the sandwich format outlined in Examples 1 and 2.
  • smaller analytes such as drugs, including therapeutic drugs and illicit drugs, hormones, vitamins, steroids, peptides and haptens as well as the larger molecular weight molecules which can be assayed in the sandwich format outlined in Examples 1 and 2.
  • FIG. 8 A side view diagrammatic representation of a competition assay in accordance with the invention is shown in FIG. 8.
  • the device consists of a filter 601 adapted to remove red blood cells, a separate support element 602 containing a suitable concentration of reagents including anti- digoxin gold-labeled antibody 603 (i.e., analyte detection agent) specific for the drug digoxin 611 (i.e. target analyte) and a streptavidin-gold 604 calibration agent.
  • anti- digoxin gold-labeled antibody 603 i.e., analyte detection agent
  • streptavidin-gold 604 calibration agent i.e., streptavidin-gold 604 calibration agent
  • the test device comprises a nitrocellulose membrane 605 upon which is non-diffusibly attached at first and second calibration zones 606, 608 different predetermined amounts 614, 615 of BSA-biotin 604 which when in operation generate different signal intensities (FIG. 8).
  • the membrane 605 includes a test zone 607 whereat there is non-diffusibly attached an unlabeled digoxin (or an unlabeled analog thereof) 613 which is capable of binding specifically to the anti-digoxin gold-labeled antibody 603.
  • the test zone has non- diffusibly attached therein an unlabeled anti-digoxin antibody which is capable of binding specifically the labeled digoxin and unlabeled digoxin if present in the test sample.
  • the membrane also includes a procedural control zone 609 which has non-diffusibly attached therein an anti-mouse antibody 616 capable of binding the anti-digoxin gold- labeled antibody 603 which has not bound to the test zone 607. Adjacent to one end of the membrane 605, there is also provided an absorbent pad 610 which absorbs an excess amount of the test sample, the application buffer or assay reagents. It will also be appreciated that when labeled digoxin is employed as the analyte detection agent in, for example a competitive assay, an anti-digoxin antibody may be used as a capturing component in the procedural control zone.
  • test sample and/or the application buffer causes moistening of the separate support element 602 and subsequent mobilization of the anti-digoxin gold-labeled antibody 603 and the streptavidin-gold 604.
  • digoxin 611 is present in the test sample, it binds and complexes with the anti-digoxin gold-labeled antibody 603 and, in concert with the streptavidin-gold 604, migrates subsequently to a detection area comprising the test zone 607 and calibration zones 606, 608.
  • the binding of anti-digoxin gold-labeled antibody 603 to the attached unlabeled digoxin 613 is inhibited in the presence of digoxin 611. This results in a test zone 607 of reduced intensity compared to that generated in the absence of digoxin 611.
  • a zone of strong intensity is generated in the test zone 607.
  • the digoxin if present in the test sample, and the labeled digoxin migrate along the membrane without significant interaction. In the test zone, the binding of labeled digoxin to non-diffusibly attached unlabeled anti-digoxin antibody is inhibited in the presence of digoxin.
  • the concentrations ofthe streptavidin-gold 604 and BSA-biotin 612 are adjusted to give signal intensities that fall within a suitable dynamic range of those produced by typical therapeutic concentrations of digoxin 611 as described in Example 6.
  • the lower calibration zone when in use, generates signal intensities corresponding to approximately 0.8 ng/mL of digoxin.
  • the limit of detection of this analyte in a flow through device is 0.4 ng/mL and consequently, the use of calibration zones generating signal intensities corresponding to 0.8 ng/mL and 1.6 ng/mL respectively enable visual inspection of a test device to assign one of four values to a given sample (FIG. 9).
  • FIG. 9a there is shown a top view of an unused device for semi-quantitative measurement of digoxin. Notice that there are no visible instrumental read lines in the viewing window comprising procedural control (C), test (T), 0.8 ng/mL and 1.6 ng/mL calibration zones.
  • FIG. 9b there is shown the test device after the successful completion of a test not containing sub-therapeutic doses of digoxin. Note the appearance of zones of color in the 0.8 ng/mL and 1.6 ng/mL calibration zones and the procedural control zone and the intense color generated in the test zone. This allows a value of ⁇ 0.8 ng/mL digoxin to be assigned to this test sample.
  • FIG. 9c there is shown the device after the successful completion of test with a sample containing between 0.8 ng/mL to 1.6 ng/mL digoxin.
  • a visible line results in the test zone and is of lower intensity than that generated in the 0.8 ng/mL calibration zone but stronger than that generated in the 1.6 ng/mL calibration zone allowing a value of between 0.8-1.6 ng/mL of digoxin (the normal therapeutic range) to be assigned to this sample.
  • the signal intensities of the other zones are similar to those shown in FIG. 9b.
  • FIG. 9d there is shown the device after the successful completion of a test on a sample containing a digoxin concentration of > 1.6 ng/mL (toxic range). Note the zone of intensity in the viewing window; the test zone has generated a signal intensity lower than that generated in the 1.6 ng/mL calibration zones allowing a higher value to be assigned to the sample. Any unbound labeled reagents migrate to the absorbent pad where they are not visible to the observer.
  • FIG. 10 shows a side view diagrammatic representation ofthe assay reagents after completion of a test in which no digoxin was present in the test sample.
  • the labeled reagents 603, 604 have migrated from the separate support element 602 and the streptavidin-gold has bound to the BSA-biotin 612 in the calibration zones 606, 608.
  • the anti-digoxin gold- labeled antibody 603 has bound to the attached unlabeled digoxin 613 in the test zone 607 generating a maximal signal.
  • FIG. 11 shows a completed test in which toxic concentrations of digoxin 611 were present in the test sample.
  • the streptavidin- gold has been captured at calibration zones 606, 608 but the digoxin 611 has inhibited the binding of the anti-digoxin gold-labeled antibody 603 at the test zone generating a reduced signal compared to that generated in the absence of digoxin 611 in the test sample.
  • the lateral flow liquid permeable medium in accordance with the invention may be read visually after completion of an assay, as described in Examples 1 to 3.
  • the lateral flow liquid permeable medium can be read in an instrument configured to receive the device and to measure the change in optical reflectivity produced by the attachment ofthe label onto the surface of the membrane.
  • the instrument is preferably configured to measure reflectance at a wavelength corresponding to the appropriate, usually maximum, absorbance of the label. It can include more than one LED or suitable light source for measuring and detecting a multiple of reflected wavelengths.
  • the instrument is programmed to contain standard curves for the various tests to be read and each test has a code, interpreted by the instrument, such that the correct data for a specific target analyte is accessed.
  • the instrument can be a stand alone instrument, which prints out or displays a result for the operator, or it can be connected to the laboratory's central computer and aligned with other patient data.
  • the results obtained in this way are fully quantitative and such an instrument is valuable in monitoring a patient's analyte levels during therapy or for following the course of disease or progress after surgery.
  • FIG. 12 A comparison of a quantitative determination of D-dimer by instrument reading with D-dimer concentration as determined by enzyme immunoassay for D-dimer is shown in FIG. 12. The results are presented as a linear regression analysis, thus, indicating a strong correlation between instrument reading and EIA.
  • This instrument reading comparison was made with 187 plasma samples of clinical origin using 25 ⁇ L of sample. At ten minutes after the tests were run the reflectance signal of the test zone was compared with the reflectance signal of the calibration zones. The instrument readings were performed using a simple reflectance instrument (Tobias IQ 150 Portable Computing Reflection Desitometer, Tobias Associates Inc, Ivyland, PA, USA).
  • FIG. 13 A comparison of a quantitative determination of myoglobin by instrument reading with the myoglobin concentration as determined by enzyme immunoassay for myoglobin is shown in FIG. 13. The results are presented as a linear regression analysis, thus, indicating a strong correlation between instrument reading and EIA.
  • This instrument reading comparison was made with 127 plasma samples of clinical origin using 25 ⁇ L of sample. At ten minutes after the tests were run the reflectance signal of the test zone was compared with the reflectance signal of the calibration zones. The instrument readings were performed using the reflectance instrument referred to in Example 6.
  • a series of test strips comprising the lateral flow liquid permeable medium of the invention are run containing a fixed concentration of calibration agent, e.g. streptavidin-gold with the strips coated at the calibration zone with a fixed BSA concentration but with a variable biotin content varying from 0.1 to 3.0 moles of biotin per mole of BSA.
  • a fixed concentration of calibration agent e.g. streptavidin-gold
  • biotin content varying from 0.1 to 3.0 moles of biotin per mole of BSA.
  • the reflectance signal at the calibration zone is determined (FIG. 14).
  • the reflectance signal is plotted against the concentration of biotin-BSA and is used as a standard curve as follows: For a given target analyte, a small number of samples with a known analyte concentration are run on strips containing target detection agent and target capture agent. The reflectance signal at the capture zone is then read off the standard curve of reflectance signal versus biotin-BSA concentration to determine the concentration of the calibration capture (biotin-BSA) which is required to match a particular clinical cut-off value (Table 3). This is of particular use in the production of tests in which a semi- quantitative determination of analyte concentration is required.
  • calibrators can be chosen which cover the dynamic range of the target analyte which is determined as described above using the reflectance signals of samples with known values run on tests containing target analyte detection and calibration agents.
  • BSA is biotinylated using NHS-LC-Biotin in carbonate buffer to yield 25 moles of biotin per mole of BSA. This stock solution of biotin-BSA is then diluted in BSA to the required concentration.
  • the labelled analyte detection agent and labelled calibration agent may be applied to the lateral flow liquid permeable medium in solution and, thus, these reagents may be included with the test sample if desired.
  • the affinity and avidity of the calibration zone reactants which may be used are significantly greater than the test zone reactants which substantially eliminates interference and facilitates quantitation of a wide range of analytes as described above.
  • the calibration zone reactions suitably have an affinity constant of 10 "15 M which is six orders of magnitude higher affinity than normal antibody affinities;
  • the assay is essentially continuous in operation and thus avoids discontinuity which is a prerequisite of multiple step assays.
  • TABLES TABLE 1 Semi-quantitative determination of D-dimer effected by comparison of respective signals at the test and calibration zones
  • a signal intensity at the test Greater than 120 ng/mL but less zone that is between the than 1000 ng/mL intensity of the two calibration zones
  • a signal intensity at the test zone that is Greater than 0.8 between the intensity of the calibration ng/mL but less than zones 1.6 ng/mL
  • Myoglobin 0.5 moles biotin/mole BSA 70 ng/mL 640
  • Canine Heartworm 0.25 moles biotin/mole BSA 100 ng/mL 275
  • FIG. 14 Standard Curve of Biotin-BSA Concentration vs Reflectance Signal.

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Abstract

Cette invention se rapporte à un procédé permettant l'évaluation semi-quantitative d'une substance cible à analyser dans un échantillon d'essai. Ledit procédé consiste (i) à attacher, d'une manière ne permettant pas la diffusion, à au moins une zone de mesure d'un milieu perméable aux liquides à flux latéral, un récepteur de la substance à analyser capable de se lier à la substance cible ou une quantité préétablie de ladite substance, (ii) à attacher, d'une manière permettant la diffusion, à un milieu de support qui peut comprendre le milieu perméable aux liquides à flux latéral ou à un élément de support distinct, un agent de détection de la substance qui détecte la présence de substance cible dans l'échantillon d'essai, ledit agent de détection de la substance étant associé à un marqueur, (iii) à attacher, d'une manière permettant la diffusion, à un milieu de support qui peut comprendre le milieu perméable aux liquides à flux latéral ou à un élément de support distinct, un agent d'étalonnage associé à un marqueur, (iv) à attacher, d'une manière ne permettant pas la diffusion, à au moins une zone d'étalonnage du milieu perméable aux liquides à flux latéral, un récepteur de l'agent d'étalonnage capable de se lier à l'agent d'étalonnage, (v) à mettre en contact le milieu perméable aux liquides à flux latéral avec l'échantillon d'essai et (vi) à comparer les signaux associés à chaque marqueur au niveau de la (des) zone(s) de mesure et de la (des) zone(s) d'étalonnage de façon à permettre l'évaluation de la substance cible dans l'échantillon d'essai.
EP96928285A 1995-09-07 1996-09-09 Procede et appareil destines a l'evaluation quantitative et semi-quantitative d'une substance a analyser Withdrawn EP0864090A4 (fr)

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AUPN5279A AUPN527995A0 (en) 1995-09-07 1995-09-07 Method and apparatus for semiquantification of an analyte
AUPN5279/95 1995-09-07
PCT/AU1996/000557 WO1997009620A1 (fr) 1995-09-07 1996-09-09 Procede et appareil destines a l'evaluation quantitative et semi-quantitative d'une substance a analyser

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Families Citing this family (53)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5935522A (en) * 1990-06-04 1999-08-10 University Of Utah Research Foundation On-line DNA analysis system with rapid thermal cycling
US5753517A (en) * 1996-03-29 1998-05-19 University Of British Columbia Quantitative immunochromatographic assays
AU6277498A (en) * 1997-02-15 1998-09-08 Beth Israel Deaconess Medical Center Multiple-site antibody capture immunoassays and kits
SE9704933D0 (sv) * 1997-12-30 1997-12-30 Pharmacia & Upjohn Diag Ab Metod som utnyttjar en ny kalibrator och test kit som innehåller kalibratorn
US7713703B1 (en) 2000-11-13 2010-05-11 Biosite, Inc. Methods for monitoring the status of assays and immunoassays
WO2000031538A1 (fr) * 1998-11-23 2000-06-02 Praxsys Biosystems, Inc. Essais en ecoulement lateral ameliores
US6136610A (en) * 1998-11-23 2000-10-24 Praxsys Biosystems, Inc. Method and apparatus for performing a lateral flow assay
FR2788856B1 (fr) * 1999-01-21 2001-07-13 Adiatec Sa Dispositif de test et procede pour la quantification de resultats obtenus par des methodes immunochromatographiques
US6551842B1 (en) * 1999-03-26 2003-04-22 Idexx Laboratories, Inc. Method and device for detecting analytes in fluids
JP2000321277A (ja) * 1999-05-13 2000-11-24 Matsushita Electric Ind Co Ltd クロマト定量測定装置
US6509196B1 (en) * 2000-01-04 2003-01-21 Response Biomedical Corp. Compensation for non-specific signals in quantitative immunoassays
JP4562854B2 (ja) * 2000-05-08 2010-10-13 パナソニック株式会社 クロマトグラフィー測定方法
JP3726082B2 (ja) * 2000-12-26 2005-12-14 松下電器産業株式会社 特異結合分析方法およびそれに用いる特異結合分析装置
EP1459068B1 (fr) * 2001-12-24 2010-08-18 Kimberly-Clark Worldwide, Inc. Syst me d' talonnage interne pour essai en coulement continu
US20030119203A1 (en) * 2001-12-24 2003-06-26 Kimberly-Clark Worldwide, Inc. Lateral flow assay devices and methods for conducting assays
US6837171B1 (en) 2002-04-29 2005-01-04 Palmer/Snyder Furniture Company Lightweight table with unitized table top
US7175992B2 (en) 2002-04-10 2007-02-13 Response Biomedical Corporation Sensitive immunochromatographic assay
JP4532121B2 (ja) 2002-04-10 2010-08-25 レスポンス バイオメディカル コーポレイション 高感度の免疫クロマトグラフィーアッセイ法
US7314763B2 (en) * 2002-08-27 2008-01-01 Kimberly-Clark Worldwide, Inc. Fluidics-based assay devices
US7285424B2 (en) 2002-08-27 2007-10-23 Kimberly-Clark Worldwide, Inc. Membrane-based assay devices
CN100473989C (zh) * 2002-08-27 2009-04-01 金伯利-克拉克环球有限公司 采用时间分辨荧光的基于膜的检测
FI20021935A (fi) * 2002-10-31 2004-05-01 Erilab Oy Menetelmä pikatestiä varten ja kontrollilaite
US7781172B2 (en) 2003-11-21 2010-08-24 Kimberly-Clark Worldwide, Inc. Method for extending the dynamic detection range of assay devices
US7247500B2 (en) 2002-12-19 2007-07-24 Kimberly-Clark Worldwide, Inc. Reduction of the hook effect in membrane-based assay devices
US20040121334A1 (en) 2002-12-19 2004-06-24 Kimberly-Clark Worldwide, Inc. Self-calibrated flow-through assay devices
FR2853077B1 (fr) * 2003-03-28 2005-12-30 Vedalab Procedes immunochromatographiques en phase solide
JP4854506B2 (ja) 2003-04-01 2012-01-18 ユニバーシティ オブ サザン カリフォルニア 疾患のリスクを予測および評価するためのう食リスク試験
US20040197819A1 (en) 2003-04-03 2004-10-07 Kimberly-Clark Worldwide, Inc. Assay devices that utilize hollow particles
US7851209B2 (en) 2003-04-03 2010-12-14 Kimberly-Clark Worldwide, Inc. Reduction of the hook effect in assay devices
US8101429B2 (en) 2003-06-03 2012-01-24 Siemens Healthcare Diagnostics Inc. Native analyte as a reference in lateral flow assays
DK1646862T3 (en) * 2003-06-03 2018-07-23 Siemens Healthcare Diagnostics Inc Automatic identification of reagent test strips using reflection values
DE10337772B4 (de) * 2003-08-14 2009-03-05 Seratec Gesellschaft für Biotechnologie mbH Immunoassay und Nachweisverfahren
US7943395B2 (en) 2003-11-21 2011-05-17 Kimberly-Clark Worldwide, Inc. Extension of the dynamic detection range of assay devices
US7713748B2 (en) 2003-11-21 2010-05-11 Kimberly-Clark Worldwide, Inc. Method of reducing the sensitivity of assay devices
US20050112703A1 (en) * 2003-11-21 2005-05-26 Kimberly-Clark Worldwide, Inc. Membrane-based lateral flow assay devices that utilize phosphorescent detection
US7943089B2 (en) 2003-12-19 2011-05-17 Kimberly-Clark Worldwide, Inc. Laminated assay devices
EP1612558A1 (fr) * 2004-06-30 2006-01-04 Academisch Ziekenhuis Leiden Procédé de dosage du gluten
US7521226B2 (en) 2004-06-30 2009-04-21 Kimberly-Clark Worldwide, Inc. One-step enzymatic and amine detection technique
AU2005259012B2 (en) * 2004-07-01 2011-01-20 Forsite Diagnostics Limited Analyte detection system
CN101031798B (zh) 2004-07-29 2012-06-27 瑞莱诊断体系有限公司 侧向流系统和测定
FR2876453B1 (fr) * 2004-10-11 2007-01-12 Biomerieux Sa Procede de diagnostic d'exclusion in vitro des syndromes coronariens aigus
CA2588230A1 (fr) 2004-11-23 2006-08-10 Response Biomedical Corporation Dosage immunologique mettant en oeuvre une reaction d'etalonnage interne en deux etapes
US7939342B2 (en) * 2005-03-30 2011-05-10 Kimberly-Clark Worldwide, Inc. Diagnostic test kits employing an internal calibration system
US20060275920A1 (en) * 2005-06-01 2006-12-07 Petrilla John F Apparatus and method for discriminating among lateral flow assay test indicators
US11506674B2 (en) * 2005-07-29 2022-11-22 Princeton Biomeditech Corporation Monoclonal antibody against D-dimer and diagnosis agent for detecting D-dimer, crosslinked fibrin and its derivatives containing D-dimer by using the antibody
US7829347B2 (en) * 2005-08-31 2010-11-09 Kimberly-Clark Worldwide, Inc. Diagnostic test kits with improved detection accuracy
GB0604973D0 (en) * 2006-03-11 2006-04-19 Central Science Lab Csl Of San Purification method and kit
WO2008075193A2 (fr) * 2006-12-19 2008-06-26 Inverness Medical Switzerland Gmbh Dispositifs et procédés de dosage
WO2008106021A1 (fr) 2007-02-26 2008-09-04 Response Biomedical Corporation Dosage comparatif de multiples analytes
CN102197307A (zh) * 2008-08-22 2011-09-21 电化生研株式会社 用于膜分析的配有参照显示部分的试验装置
EP2194381B1 (fr) 2008-12-03 2015-12-02 Roche Diagnostics GmbH Elément de test doté d'une zone de contrôle et de calibrage combinée
CN102375071B (zh) * 2011-09-21 2014-04-16 艾康生物技术(杭州)有限公司 生物样本参数的测试方法
US10082501B2 (en) 2013-03-24 2018-09-25 Biomedomics, Inc. Nanotube based lateral flow device for biomarker detection

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0314499A1 (fr) * 1987-10-30 1989-05-03 Unilever Plc Dispositifs et procédés pour des essais chimiques
EP0560411A2 (fr) * 1987-04-27 1993-09-15 Unilever N.V. Dispositif d'analyse pour l'exécution d'essais de liaisons spécifiques
EP0383619B1 (fr) * 1989-02-17 1997-04-23 Unilever Plc Dispositif analytique en phase solide

Family Cites Families (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW203120B (fr) * 1985-10-04 1993-04-01 Abbott Lab
US4868108A (en) * 1985-12-12 1989-09-19 Hygeia Sciences, Incorporated Multiple-antibody detection of antigen
JPS62231168A (ja) * 1986-03-21 1987-10-09 ハイブリテツク・インコ−ポレイテツド アナライト−レセプタ−分析用内部標準を設けるための改良法
US4960691A (en) * 1986-09-29 1990-10-02 Abbott Laboratories Chromatographic test strip for determining ligands or receptors
CA1339723C (fr) * 1988-01-19 1998-03-17 Philip Mcmahon Immunoesssai a point d'essai multiples
DE68924802T2 (de) * 1988-05-17 1996-05-23 Enzo Biochem Inc Testvorrichtung mit gradientreguliertem lokalisiertem Signal.
AU3439693A (en) * 1992-01-22 1993-09-01 Abbott Laboratories Calibration reagents for semi-quantitative binding assays and devices
FR2702841B1 (fr) * 1993-03-19 1996-01-26 Toledano Jacques Dispositif et procédé immunologique peptide-anti-peptide.

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0560411A2 (fr) * 1987-04-27 1993-09-15 Unilever N.V. Dispositif d'analyse pour l'exécution d'essais de liaisons spécifiques
EP0314499A1 (fr) * 1987-10-30 1989-05-03 Unilever Plc Dispositifs et procédés pour des essais chimiques
EP0383619B1 (fr) * 1989-02-17 1997-04-23 Unilever Plc Dispositif analytique en phase solide

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO9709620A1 *

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